Stress Estimation of Pre-Slaughter and Slaughtered by Means of Fourier Transform Infrared Spectroscopy Analysis Through Measurement of Cortisol and Catecholamine Level in Female Cattle Urine

Abstract. This study was aimed to analyze the levels of catecholamine and cortisol as stress indicator by invasive and non-invasive methods. Twelve female cattle PO were used in this study and were collected from slaughter house in Yogyakarta. Catecholamine and cortisol level of urine were measured by Enzyme linked Immunoassay (EIA) method and Fourier Transform Infrared (FTIR), were statistically analyzed to determine the difference between pre slaughter and slaughter conditions. FTIR spectra were analyzed using chemo-metrics software.  These results showed that the concentration of urinary cortisol were 2.12±1.68 ng/dl of pre-slaughter and 7.58±3.89 ng/dl of slaughtered respectively. The levels of urinary catecholamine in pre slaughter and slaughter were 3.07±2.05 ng/dl  and 4.15±2.68 ng/dl respectively. In FTIR analysis showed the spectral separation between the different quadrants before and during slaughter. The result suggested the correlation between the results of the analysis using the EIA and FTIR spectra. It is assumed that the separation of the FTIR spectrum in line with the increased levels of cortisol catecholamine and samples. It can be concluded that FTIR can be used to analyze the status of stress in animals, especially in cattle. Key words: catecholamine, cortisol, non-invasive, FTIR Abstrak. Penelitian ini bertujuan untuk menganalisis kadar katekolamin dan kortisol sebagai indicator stress secara invasif maupun non invasif dan mengembangkan metode deteksi stress pada sapi. Penelitian menggunakan sampel urin sapi Peranakan Ongole (PO) yang diambil dari rumah potong hewan di Yogyakarta. Analisis kadar katekolamin (CA) dan kortisol (CO) dilakukan dengan menggunakan metode EIA dan FTIR. Data konsentrasi kortisol maupun katekolamin hasil pengukuran menggunakan metode EIA dilakukan analisis statistik untuk mengetahui pengaruh stres terhadap konsentrasi CA dan CO urine. Hasil penelitian ini menunjukan rata-rata konsentrasi kortisol urin adalah 2.12±1.68 ng/dl dalam kondisi prapenyembelihan dan 7,58±3,89 ng/dl pada saat penyembelihan. Sedangkan konsentrasi katekolamin urin pada saat prapenyembelihan dan penyembelihan secara berurutan adalah 3,07±2,05 ng/dl  dan 4,15±2,68 ng/dl. Analisis FTIR menggambarkan pemisahan spektral pada quadran berbeda antara sebelum dan saat penyembelihan. Berdasarkan hasil ini menunjukkan korelasi antara hasil analisis dengan menggunakan EIA dan FTIR, baik konsentrasi katekolamin dan kortisol dengan spektra FTIR dari sampel. Perbedaan konsentrasi CA dan CO urin prapenyembelihan dan saat penyembelihan sejelan dengan pemisahan spektra FTIR. Hal ini diyakini bahwa pemisahan spektrum FTIR disebabkan adanya peningkatan kadar katekolamin dan kortisol dari sampel. Dapat disimpulkan bahwa FTIR dapat untuk menganalisis status stres pada hewan khususnya pada sapi. Kata kunci : katekolamin, kortisol, non invasif, FTI

Study of advanced rheumatoid arthritis

INTRODUCTION: Rheumatic diseases are considered public health problems affecting millions of people worldwide resulting in high and rising health-care costs. In this work, Fourier Transform Infrared spectroscopy associated to Partial Least Square regression (PLS) analysis was used to diagnose rheumatoid arthritis (RA) from human serum. METHODS: The sera of 94 individuals were collected, which included 47 from rheumatic patients and 47 from healthy individuals. The results from PLS analysis were compared to standard clinical trials such as anti-citrullinated peptide antibodies, C- Reactive protein, and Rheumatoid factor. RESULTS: For clinical diagnosis, the anti-citrullinated peptide antibodies of second generation proved to be the most specific to diagnosis rheumatoid arthritis even after long periods of drug therapy. CONCLUSIONS: The qualitative PLS analysis has shown higher values of IgM of RA group, but the difference was very small. The RA patients were under medication, which interfered with the IgM concentration.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade do Vale do Paraíba Instituto de Pesquisa e DesenvolvimentoUniversidade Federal de São Paulo (UNIFESP) Departamento de Medicina Divisão de ReumatologiaUNIFESP, Depto. de Medicina Divisão de ReumatologiaSciEL

Investigations into the Effects of pH on Quantitative Measurements of Lactate in Biological Media Using ATR-FTIR Spectroscopy

Quantification of lactate/lactic acid in critical care environments is essential as lactate serves as an important biochemical marker for the adequacy of the haemodynamic circulation in shock and of cell respiration at the onset of sepsis/septic shock. Hence, in this study, ATR-FTIR was explored as a potential tool for lactate measurement, as the current techniques depend on sample preparation and fails to provide rapid response. Moreover, the effects of pH on PBS samples (7.4, 7, 6.5 and 6) and change in solution conditions (PBS to whole blood) on spectral features were also investigated. A total 189 spectra from five sets of lactate containing media were obtained. Results suggests that lactate could be measured with more than 90% accuracy in the wavenumber range of 1500-600 cm-1. The findings of this study further suggest that there exist no effects of change in pH or media, when estimating lactate concentration changes in this range of the Mid-IR spectral region

The efficacy of support vector machines in modelling deviations from the Beer-Lambert law for optical measurement of lactate

Lactate is an important biomarker with a significant diagnostic and prognostic ability in relation to life-threatening conditions and diseases such as sepsis, diabetes, cancer, pulmonary and kidney diseases, to name a few. The gold standard method for the measurement of lactate relies on blood sampling, which due to its invasive nature, limits the ability of clinicians in frequent monitoring of patients' lactate levels. Evidence suggests that the optical measurement of lactate holds promise as an alternative to blood sampling. However, achieving this aim requires better understanding of the optical behavior of lactate. The present study investigates the potential deviations of absorbance from the Beer-Lambert law in high concentrations of lactate. To this end, a number of nonlinear models namely support vector machines with quadratic, cubic and quartic kernels and radial basis function kernel are compared with the linear principal component regression and linear support vector machine. Interestingly, it is shown that even in extremely high concentrations of lactate (600 mmol/L) in a phosphate buffer solution, the linear models surpass the performance of the other models

Pre-slaughter Stress Estimation by Fourier Transform Infrared Spectroscopy Analysis

The objective of the study was to analyze the potential of fourier transform infrared (FTIR) asstress estimation instrument through measurement of catecholamine (CA) and cortisol (CO) levels byinvasive and non-invasive methods. Twelve heads of Ongole grade (PO) cow from slaughterhouse wereused in this study. Twenty four urine samples of pre-slaughter and slaughtering were collected toevaluate the CA and CO levels by ELISA method and Fourier Transform Infrared (FTIR). Data of CAand CO levels were statistically analyzed to determine the difference between pre slaughter andslaughter conditions. FTIR spectra were analyzed using chemo-metrics software. The results showedthat the levels of urinary cortisol were 2.12±1.68 ng/dl of pre-slaughter and 7.58±3.89 ng/dl ofslaughtered and the levels of urinary catecholamine in pre slaughter and slaughter were 3.07±2.05 and4.15±2.68 ng/dl respectively. The results showed a correlation between the FTIR spectra and the resultsof ELISA analysis. FTIR spectrums were distributed in different quadrants, this was caused bydifferences of CA and CO levels between pre-slaughter and slaughtered. It can be concluded that FTIRcan be supposed to analyze the status of stress in animals, especially in cattle

Imagerie IRTF tridimensionnelle pour l'étude de l'insuffisance rénale chronique

CKD (Chronic Kidney Disease) is one of the worst public diseases in developing countries. The stages of CKD are mainly based on measured or estimated GFR (Glomerular Filtration Rate). However, this method is not sensitive enough on early stages of the pathology and thus do not offer accurate diagnostic value. Early detection and treatment can often limit or avoid the chronicity effects of the disease. This thesis focuses on the development of FTIR microscopy as a diagnostic tool for the identification by histopathology at glomerulus level of the kidney in CKD model. We developed a technique of 3D reconstruction for the FTIR imaging of biochemical components changes in glomeruli for identifying the pathological marker of CKD. The curve-fitting and spectral clustering are applied on the FTIR microscopy analysis to distinguish between healthy and pathological glomeruli of a kidney. Then, the glomerular microvasculatureis highlighted to reveal the morphological abnormalities by perfusing contrast agents into blood vessels. With advanced 3D statistical methods and 3D image visualization by microscopy, FTIR spectro-imaging can be used as a functional technique to determine the morphological and molecular changes occurring along CKD development.L’insuffisance rénale chronique (IRC) et l’une des pires maladies chroniques dans les pays développés. Les grades de l’IRC sont principalement basés sur la mesure ou l’estimation du taux de filtration rénale (GFR). Cependant, cette méthode est peu sensible sur les premiers stades de la pathologie et n’apporte donc pas de valeur diagnostique. La détection de la pathologie à des stades précoces et son traitement peuvent éviter ou limiter les effets délétères de la chronicité. Cette thèse se penche sur le développement de la microscopie IRTF en tant qu’outil diagnostic pour l’identification par histopathologie à l’échelle du glomérule dans un modèle d’IRC. Nous avons développé la technique de reconstruction 3D pour l’imagerie IRTF des modifications biochimiques à l’échelle du glomérule pour déterminer des marqueurs de l’IRC. La déconvolution spectrale et le clustering sont appliqués après analyses IRTF pour distinguer les modèles sains et pathologiques. Ensuite, la microvasculature glomérulaire est révélée par agent de contraste pour en déterminer les anomalies morphologiques. Grâce aux résultats obtenus en 3D et l’utilisation de méthodes statistiques avancées, la microscopie IRTF est utilisée comme une technique fonctionnelle pour déterminer les modifications morphologiques et moléculaires apparaissant au cours du développement de l’IRC

Comparison of wavelength selection methods for in-vitro estimation of lactate: a new unconstrained, genetic algorithm-based wavelength selection

Biochemical and medical literature establish lactate as a fundamental biomarker that can shed light on the energy consumption dynamics of the body at cellular and physiological levels. It is therefore, not surprising that it has been linked to many critical conditions ranging from the morbidity and mortality of critically ill patients to the diagnosis and prognosis of acute ischemic stroke, septic shock, lung injuries, insulin resistance in diabetic patients, and cancer. Currently, the gold standard for the measurement of lactate requires blood sampling. The invasive and costly nature of this procedure severely limits its application outside intensive care units. Optical sensors can provide a non-invasive, inexpensive, easy-to-use, continuous alternative to blood sampling. Previous efforts to achieve this have shown significant potential, but have been inconclusive. A measure that has been previously overlooked in this context, is the use of variable selection methods to identify regions of the optical spectrum that are most sensitive to and representative of the concentration of lactate. In this study, several wavelength selection methods are investigated and a new genetic algorithm-based wavelength selection method is proposed. This study shows that the development of more accurate and parsimonious models for optical estimation of lactate is possible. Unlike many existing methods, the proposed method does not impose additional locality constraints on the spectral features and therefore helps provide a much more granular interpretation of wavelength importance

Utilisation de la spectro-imagerie IR-TF pour le développement d'une anatomo-pathologie moléculaire des tumeurs cérébrales

Les gliomes sont des tumeurs agressives de mauvais pronostic, très angiogéniques et infiltrantes ce qui rend leur exérèse particulièrement difficile. Vu les limites des techniques actuelles d imagerie, nous avons proposé la spectro-imagerie Infrarouge à Transformée de Fourier (IRTF), d une résolution spatiale de 6 m, pour apporter une information moléculaire à l examen histologique actuel des gliomes. Nos travaux ont été fondés sur la recherche de paramètres moléculaires des vaisseaux sanguins, notamment sur la base des contenus de leur membrane basale. Celle-ci subit des altérations dûes au stress angiogénique tumoral. Nous avons mis en évidence des altérations de la structure secondaire des protéines (tels les collagènes) des vaisseaux sanguins au cours de la croissance de la tumeur. Nous avons aussi évalué les modifications des chaines d acides gras des phospholipides membranaires, qui révélent un degré d insaturation plus important pour les vaisseaux tumoraux. Ensuite, sur un modèle de gliome murin, nous avons établi une méthode efficace de classification des capillaires sanguins sur la base d absorptions de leurs contenus glucidiques et lipidiques, permettant de discriminer totalement les capillaires sains et tumoraux. La combinaison de ces paramètres a été mise à profit pour assurer une histopathologie moléculaire des gliomes humains. Nos résultats ont démontré qu il est possible de différencier entre la vasculature saine et tumorale sur ces gliomes humains, ce qui permet une bonne délimitation des zones tissulaires correspondantes. Cette technique pourrait devenir un outil analytique fiable, rapide d une durée compatible avec la chirurgie et donc très utile pour les neurochirurgiens.Malignant gliomas are very aggressive tumors with poor prognosis, highly angiogenic and invasive into the surrounding brain parenchyma, making their resection very difficult. Regarding the limits of current imaging techniques, we have proposed Fourier Transform Infrared (FTIR) spectro-imaging, with a spatial resolution of 6 m, to provide molecular information for the histological examination of gliomas. Our work was based on the research of molecular parameters of blood vessels, notably on the basis of the contents of their basement membrane, which undergoes changes due to tumor angiogenic stress. We have identified alterations of the secondary structure of proteins (such as collagen) in blood vessels during tumor growth. We have also assessed the changes in fatty acyl chains of membrane phospholipids, which revealed a higher unsaturation level in tumor vessels. Then, on a murine glioma model, we have established an efficient method of blood vessels classification based on their carbohydrates and fats contents, allowing the differentiation between healthy and tumor blood vessels. The combination of these parameters was used to provide a molecular histopathology for the study of human gliomas. Our results have demonstrated the feasibility of differentiating between healthy and tumor vasculature in these human gliomas, which help delimitating areas of corresponding tissue. This technique could become a reliable and fast analytical tool, with duration compatible with the surgery and thus very useful for neurosurgeons.BORDEAUX1-Bib.electronique (335229901) / SudocSudocFranceF